Recovery solution

Manufactured by BD

Sourced in United States

Recovery Solution is a laboratory reagent designed to facilitate the recovery and purification of samples. Its core function is to provide a controlled environment for the separation and extraction of target analytes from complex matrices. The solution is formulated to maintain the integrity and stability of the samples during processing.

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Lab products found in correlation

Anti cd31 alexafluor 647, by BD (1 mentions) Matrigel, by Corning (1 mentions) Cell counter, by Beckman Coulter (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Onetouch ultra2, by LifeScan (1 mentions)

Close spelling variants of this product

Cell Recovery Solution Matrigel recovery solution

7 protocols using recovery solution

Viability and Endothelial Cell Characterization

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For viability assay, cultures were harvested on ice and resuspended in PBS containing 3% FCS and 5 mM EDTA. DAPI staining (Miltenyi) was used at 1/1,000 dilutions 5 min before analysis. For 3D recovery and viability testing, cells were dissociated from recovered spheroids using the BD Recovery Solution (BD Biosciences) following manufacturer instructions.
For endothelial cells characterization an anti-CD31-Alexafluor 647 (BD Biosciences) was used at 1/100 dilution.
For intracellular staining, cells were fixed and permeabilized using a BD Bioscience Fix/Perm assay following manufacturer instructions. Anti-SRC, phospho-SRC and FGF-2 unlabeled primary antibodies (Cell Signaling) were used 30 min at 1/100 dilution and secondary anti rabbit-Alexa Fluor 488 (Cell Signaling) was used at 1/300 for 30 min. A MACSQuant instrument was used to perform experiments and FlowJo 10.4 (Treestar Inc.) software was used to analyze all data.

Cattin S., Ramont L, & Rüegg C. (2018). Characterization and In Vivo Validation of a Three-Dimensional Multi-Cellular Culture Model to Study Heterotypic Interactions in Colorectal Cancer Cell Growth, Invasion and Metastasis. Frontiers in Bioengineering and Biotechnology, 6, 97.

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Mammary Epithelial Cell Fractionation and 3D Culturing

Cited 1 time

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Whole tissue, LEC and MEC cell fractions were prepared from mammary glands and lysed to obtain purified cell fractions (Macias et al. 2011 (link)). For preparation of single-cell suspensions for fluorescence-activated cell sorting (FACS), thoracic and inguinal mammary glands were harvested and mammary epithelial single-cell suspensions were prepared as previously described (Harburg et al. 2014 (link)). All cells for the limiting dilution analysis and basal colony 3-D Matrigel cultures were generated from FACS-purified Lin⁻CD24⁺CD29^hi (basal) cells. To stain colonies, FACS-purified basal cells were resuspended in 100% Matrigel (Corning) and 5000 cells were plated per 20 μL Matrigel in 8-well chamber slides (Labtek). Colony media (DMEM-F12, 1% FCS, 0.5 μg/mL Hydrocortisone, 1 μg/mL Insulin, 10 ng/mL EGF, 20 ng/mL Cholera toxin, 1% Pen/Strep) was added after Matrigel had solidified, and colonies grown at 37°C in hypoxic conditions. After 7 days in culture, colonies were either paraffin embedded and immunostained or counted, harvested from Matrigel using BD Recovery Solution (BD), dissociated using 0.05% Trypsin-EDTA, counted and re-plated (for PKH26 and colony passaging assays) (Harburg et al. 2014 (link)). Colony counts and diameter measurements were performed using Fiji.

Ballard M.S., Zhu A., Iwai N., Stensrud M., Mapps A., Postiglione M.P., Knoblich J.A, & Hinck L. (2015). Mammary stem cell self-renewal is regulated by Slit2/Robo1 signaling through Snail and mInscuteable. Cell reports, 13(2), 290-301.

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Clonogenic Assay for 2D and 3D Cell Culture

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For 2D culture, irradiated cells were washed with PBS buffer, trypsinized and counted using a cell counter (Coulter) after irradiation. An appropriate number of cells were plated into each 60-mm dish to produce colonies. For 3D culture, the irradiated and control cells were first recovered from Matrigel by using Recovery Solution (BD) on ice for 30 min, and then trypsinized and resuspended in medium. An appropriate number of cells were plated into each 60-mm dish to produce colonies. After incubating for 10 days, the cells were fixed with 10 mL fresh Carnoy's fluid, stained with 0.5% crystal violet for 20 min. The number of colonies with greater than 50 cells were counted as survivors. Plating efficiencies (PE) were calculated as follows: numbers of colonies formed/numbers of cells plated. Surviving fractions were calculated as follows: PE (irradiated)/PE (unirradiated). The parameters of the survival curve, such as the α and β values, were obtained from the survival fraction (SF) data by curve fitting using the nonlinear model as follows: SF = (1 + exp(−α*(D - β)))⁻¹. Where D is the radiation dose delivered to the cells. All experiments were performed in triplicate. The experiment was at least repeated for three times independently.

Pan D., Chen Y., Du Y., Ren Z., Li X, & Hu B. (2016). Methylation of promoter of RBL1 enhances the radioresistance of three dimensional cultured carcinoma cells. Oncotarget, 8(3), 4422-4435.

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Recovering 3D Structures from Matrigel

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3D structures were recovered from Matrigel using Recovery Solution (BD, USA) according to the manufacturer's instructions as described previously [17 (link), 58 (link)]. Briefly, 3D cultures were first washed with ice-cold PBS, and then the Matrigel containing the 3D structures was removed from the well, transferred to 15 mL tube containing the pre-chilled Recovery Solution (1 mL per well), incubated on ice for 45min with intermittent mixing and then centrifuged at 1000 rpm for 10 min at 4°C. The supernatant containing the dissolved Matrigel was discarded and the 3D structures were washed once with PBS. To make single-cell suspension of recovered 3D structures, cells were trypsinized using trypsin-EDTA (0.25%, Invitrogen).

Pan D., Chen Y., Du Y., Ren Z., Li X, & Hu B. (2016). Methylation of promoter of RBL1 enhances the radioresistance of three dimensional cultured carcinoma cells. Oncotarget, 8(3), 4422-4435.

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Clonogenic Assay for 2D and 3D Cell Cultures

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For 2D culture, cells were trypsinized after radiation and resuspended in medium. An appropriate number of cells were plated into each 60-mm dish to produce colonies. For 3D culture, the irradiated and control cells were first recovered from matrigel by using Recovery Solution (BD, USA) on ice for 30 min, and then trypsinized and resuspended in medium. An appropriate number of cells were plated into each 60-mm dish to produce colonies. After incubating for 10 days, both 2D- and 3D-grown cells were stained with 0.5% crystal violet for 20 min. Colonies containing >50 cells were counted as survivors. Plating efficiencies (PE) were calculated as follows: numbers of colonies formed/numbers of cells plated. Surviving fractions were calculated as follows: PE (irradiated)/PE (unirradiated).

Xue G., Ren Z., Grabham P.W., Chen Y., Zhu J., Du Y., Pan D., Li X, & Hu B. (2015). Reprogramming mediated radio-resistance of 3D-grown cancer cells. Journal of Radiation Research, 56(4), 656-662.

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Streptozotocin-Induced Diabetes Reversal

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Primary Endocrine/Acinar colonies were handpicked, pooled, and incubated in recovery solution (BD Bioscience) for 1 h on ice. Intact colonies released from the semisolid media were washed and collected for transplantation. NOD-SCID mice (8 week-old males) were injected with streptozotocin (STZ; 50 mg/kg body weight; freshly made in 0.05 M citrate buffer, pH 4.5) for 3 consecutive days to induce diabetes [41 (link)]. Hyperglycemia was defined as >200 mg/dl and measured by OneTouch Ultra2 glucometer (LifeScan, Milpitas, CA, USA) starting 1 week after the last STZ injection. Approximately 12,000 Endocrine/Acinar colonies were placed under the renal capsule per mouse (n=2). Grafts were examined 5 weeks post-transplantation by immunohistochemical staining of formalin-fixed paraffin-embedded tissue sections.

Jin L., Gao D., Feng T., Tremblay J.R., Ghazalli N., Luo A., Rawson J., Quijano J., Chai J., Wedeken L., Hsu J., LeBon J., Walker S., Shih H.P., Mahdavi A., Tirrell D.A., Riggs A.D, & Ku H.T. (2015). Cells with Surface Expression of CD133highCD71low are Enriched for Tripotent Colony-Forming Progenitor Cells in the Adult Murine Pancreas. Stem cell research, 16(1), 40-53.

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Matrigel-based 3D Cell Recovery

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Three-dimensional cultured cells were recovered from matrigel using recovery solution (BD) according to the manufacturer’s instructions as described previously (Lee et al., 2007 (link)). In brief, matrigel containing the 3D structures was first washed with ice-cold PBS and then removed from the well. After being transferred to a 15-ml tube containing the pre-chilled recovery solution (1 ml per well), the mixture was incubated on ice for 45 min with intermittent mixing and then centrifuged at 1,000 rpm for 10 min at 4°C. The supernatant containing the dissolved matrigel was discarded, and the 3D structures were washed once with PBS. To make a single-cell suspension of recovered 3D structures, cells were trypsinized using trypsin- ethylenediaminetetraacetic acid (0.25%, Invitrogen). Dissociated cells were used for colony formation assay.

Pan D., Du Y., Li R., Shen A., Liu X., Li C, & Hu B. (2021). miR-29b-3p Increases Radiosensitivity in Stemness Cancer Cells via Modulating Oncogenes Axis. Frontiers in Cell and Developmental Biology, 9, 741074.

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